Rapid prototyping enzyme homologs to improve titer of nicotinamide mononucleotide using a strategy combining cell-free protein synthesis with split GFP.

Engineering biological systems to test new pathway variants containing different enzyme homologs is laborious and time-consuming. To tackle this challenge, a strategy was developed for rapidly prototyping enzyme homologs by combining cell-free protein synthesis (CFPS) with split green fluorescent protein (GFP). This strategy featured two main advantages: (1) dozens of enzyme homologs were parallelly produced by CFPS within hours, and (2) the expression level and activity of each homolog was determined simultaneously by using the split GFP assay. As a model, this strategy was applied to optimize a 3-step pathway for nicotinamide mononucleotide (NMN) synthesis. Ten enzyme homologs from different organisms were selected for each step. Here, the most productive homolog of each step was identified within 24 h rather than weeks or months. Finally, the titer of NMN was increased to 1213 mg/L by improving physiochemical conditions, tuning enzyme ratios and cofactor concentrations, and decreasing the feedback inhibition, which was a more than 12-fold improvement over the initial setup. This strategy would provide a promising way to accelerate design-build-test cycles for metabolic engineering to improve the production of desired products.

Novel indazole skeleton derivatives containing 1,2,3-triazole as potential anti-prostate cancer drugs.

In this study, a novel batch of indazole containing 1,2,3-triazole agents were designed and synthesized. The antiproliferative activity of target compounds in four human cancer cells, PC-3 (human prostate cancer cell), MCF-7 (human breast cancer cell), HepG-2 (human hepatoma cell) and MGC-803 (human gastric cancer cell), was evaluated by thiazole blue (MTT). In the antiproliferative activity screening, we were surprised to find that most compounds have specific cytotoxicity to PC-3 cancer cells. In particular, 9a has an IC50 value of 4.42 ± 0.06 µmol/L against PC-3 cell. Cloning experiments showed that 9a could inhibit the formation of PC-3 cancer cell clone in a dose-dependent manner. Through cell cycle arrest experiment, we found that compound 9a can block the cell cycle in G2/M phase and inhibit cell proliferation. Finally, by evaluating the safety of compound 9a, we noticed that it showed fairly good safety both in vivo and in vitro. Overall, based on the biological activity evaluation and safety, analogue 9a can be viewed as a potential lead compound for further development of novel anti-prostate cancer drug.

An Automatic Head Surface Temperature Extraction Method for Top-View Thermal Image with Individual Broiler.

Surface temperature variation in a broiler's head can be used as an indicator of its health status. Surface temperatures in the existing thermograph based animal health assessment studies were mostly obtained manually. 2185 thermal images, each of which had an individual broiler, were captured from 20 broilers. Where 15 broilers served as the experimental group, they were injected with 0.1mL of pasteurella inoculum. The rest, 5 broilers, served as the control group. An algorithm was developed to extract head surface temperature automatically from the top-view broiler thermal image. Adaptive K-means clustering and ellipse fitting were applied to locate the broiler's head region. The maximum temperature inside the head region was extracted as the head surface temperature. The developed algorithm was tested in Matlab® (R2016a) and the testing results indicated that the head region in 92.77% of the broiler thermal images could be located correctly. The maximum error of the extracted head surface temperatures was not greater than 0.1 °C. Different trend features were observed in the smoothed head surface temperature time series of the broilers in experimental and control groups. Head surface temperature extracted by the presented algorithm lays a foundation for the development of an automatic system for febrile broiler identification.

Fibroblast growth factor 21 attenuates hepatic fibrogenesis through TGF-ß/smad2/3 and NF-κB signaling pathways.

Fibroblast growth factor 21 (FGF-21) is a secreted protein, which has anti-diabetic and lipocaic effects, but its ability to protect against hepatic fibrosis has not been studied. In this study, we investigated the ability of FGF-21 to attenuate dimethylnitrosamine (DMN)-induced hepatic fibrogenesis in mice and the mechanism of its action. Hepatic fibrosis was induced by injection of DMN, FGF-21 was administered to the mice once daily in association with DMN injection till the end of the experiment. Histopathological examination, tissue 4-hydroxyproline content and expressions of smooth muscle α-actin (α-SMA) and collagen I were measured to assess hepatic fibrosis. Ethanol/PDGF-BB-activated hepatic stellate cells (HSCs) were used to understand the mechanisms of FGF-21 inhibited hepatic fibrogenesis. Results showed that FGF-21 treatment attenuated hepatic fibrogenesis and was associated with a significant decrease in intrahepatic fibrogenesis, 4-hydroxyproline accumulation, α-SMA expression and collagen I deposition. FGF-21 treatment inhibited the activation of HSCs via down-regulating the expression of TGF-ß, NF-κB nuclear translocation, phosphorylation levels of smad2/3 and IκBα. Besides, FGF-21 treatment caused activated HSC apoptosis with increasing expression of Caspase-3, and decreased the ratio of Bcl-2 to Bax. In conclusion, FGF-21 attenuates hepatic fibrogenesis and inhibits the activation of HSC warranting the use of FGF-21 as a potential therapeutic agent in the treatment of hepatic fibrosis.

Fibroblast growth factor (FGF21) protects mouse liver against D-galactose-induced oxidative stress and apoptosis via activating Nrf2 and PI3K/Akt pathways.

FGF21 is recently discovered with pleiotropic effects on glucose and lipid metabolism. However, the potential protective effect of FGF21 against D-gal-induced injury in the liver has not been demonstrated. The aim of this study is to investigate the pathophysiological role of FGF21 on hepatic oxidative injury and apoptosis in mice induced by D-gal. The 3-month-old Kunming mice were subcutaneously injected with D-gal (180 mg kg(-1) d(-1)) for 8 weeks and administered simultaneously with FGF21 (5 or 1 mg kg(-1) d(-1)). Our results showed that the administration of FGF21 significantly alleviated histological lesion including structure damage, degeneration, and necrosis of hepatocytes induced by D-gal, and attenuated the elevation of liver injury markers, serum AST, and ALP in a dose-dependent manner. FGF21 treatment also suppressed D-gal-induced profound elevation of ROS production and oxidative stress, as evidenced by an increase of the MDA level and depletion of the intracellular GSH level in the liver, and restored the activities of antioxidant enzymes SOD, CAT, GSH-Px, and T-AOC. Moreover, FGF21 treatment increased the nuclear abundance of Nrf2 and subsequent up regulation of several antioxidant genes. Furthermore, a TUNEL assay showed that D-gal-induced apoptosis in the mouse liver was significantly inhibited by FGF21. The expression of caspase-3 was markedly inhibited by the treatment of FGF21 in the liver of D-gal-treated mice. The levels of PI3K and PBK/Akt were also largely enhanced, which in turn inactivated pro-apoptotic signaling events, restoring the balance between pro- and anti-apoptotic Bcl-2 and Bax proteins in the liver of D-gal-treated mice. In conclusion, these results suggest that FGF21 protects the mouse liver against D-gal-induced hepatocyte oxidative stress via enhancing Nrf2-mediated antioxidant capacity and apoptosis via activating PI3K/Akt pathway.

Long-Term Administration of Fibroblast Growth Factor 21 Prevents Chemically-Induced Hepatocarcinogenesis in Mice.

PURPOSE: In this study, we explored whether treatment with FGF-21 could prevent diethylnitrosamine (DEN) induced hepatocarcinogenesis in mice. METHODS & RESULTS: Hepatoma was induced by injection of DEN every three days for 18 weeks. For the prophylactic experiment, mice were firstly injected with FGF-21 for 2 weeks, then FGF-21 was administered to the mice once daily in association with DEN injection till the end of the experiment. The hepatoma incidence of mice treated with FGF-21 was 13.3%, while the incidence of mice treated with saline was 61.5%. To understand the mechanisms, we compared the expression of ßklotho (KLB) and oxidative stress level in the livers between the mice treated with FGF-21 and saline. We found that FGF-21 could suppress DEN-induced oxidative stress and up-regulate the expression of KLB in the livers. To confirm these results, we compared the expression of KLB in L02 cells stimulated with or without FGF-21. Besides, we established DEN-induced oxidative stress cell model to affirm the relationship between FGF-21 and DEN-induced oxidative stress in vitro. Results showed that FGF-21 increased the expression of KLB and diminished the DEN-induced oxidative stress in vitro in a dose dependent manner. CONCLUSION: Systemic administration of FGF-21 can prevent DEN-induced hepatocarcinogenesis via suppressing oxidative stress and increasing the expression of KLB.

[Mechanisms of the role of fibroblast growth factor 21 in attenuating insulin resistance].

This study is to evaluate the therapeutic effect of fibroblast growth factor 21 (FGF21) on type 2 diabetic mice model and to provide mechanistic insights into its therapeutic effect. Type 2 diabetic animal model was established with high calorie fat diet and low dose streptozotocin (STZ) injection. Mice were then randomized into 5 groups: model control, FGF21 0.25 and 0.05 µmol x kg(-1) x d(-1) groups, insulin treatment group. Ten age-matched normal KM mouse administered with saline were used as normal controls. Serum glucose, insulin, lipid products and the change of serum and liver tissue inflammation factor levels between five groups of mouse were determined. The results showed that blood glucose, insulin, free fatty acids (FFAs), triglycerides, and inflammatory factor average FGF-21 of type 2 diabetes model group and normal control group were significantly higher (P < 0.01), while compared with insulin group, no difference was significant. Average blood glucose, insulin, blood lipid and inflammatory factor of FGF-21 treatment group compared with type 2 diabetes group was significantly lower (P < 0.01) and insulin group has no difference with the model control group. The results of OGTT and HOMA-IR showed that insulin resistance state was significantly relieved in a dose-dependent manner. Thus, this study demonstrates that FGF-21 significantly remits type 2 diabetic mice model's insulin resistance state and participates in the regulation of inflammatory factor levels and type 2 diabetes metabolic disorders.

[Effects of telmisartan on IKCa1 potassium channel after T-lymphocyte activation and proliferation in peripheral blood of hypertensive patients in Xinjiang Kazakh].

OBJECTIVE: To explore the effects of Telmisartan on IKCa1 potassium channel after T-lymphocyte activation and proliferation in peripheral blood of hypertensive patients in Xinjiang Kazakh. METHODS: Peripheral blood T cells in vitro culture were isolated from 30 Xinjiang Kazakh outpatients without antihypertensive drug therapy. They were randomly selected from our hypertension clinic from August 2012 to December 2012. The proliferated T lymphocytes were divided into control, telmisartan and TRAM-34 groups. After culturing for 0, 24, 48 h after corresponding treatments, the patch-clamp technique was employed to record the electrophysiological changes of IKCa1 potassium channel of T lymphocytes. RESULTS: Under different treatment conditions, the IKCa1 potassium channel showed different electrophysiological changes. Pairwise comparison was made among the groups on the same time. For the telmisartan group, IKCa1 potassium channel peak current, peak current density of intervention 24 h and 48 h were significantly reduced compared with the control group (24 h:(835 ± 117)vs(1 471 ± 255) pA, (213 ± 61) vs (388 ± 129) pA/pF; 48 h:(631 ± 142) vs (1 555 ± 383) pA, (155 ± 54) vs (388 ± 114) pA/pF, all P < 0.01) . And the blocking rates of 0 h, 24 h and 48 h of telmisartan on IKCa1 potassium channel were 6.8%, 45.1% and 60.1% respectively. CONCLUSION: Telmisartan can block the IKCa1 potassium channel of T lymphocytes in peripheral blood of hypertensive patients in Xinjiang Kazakh. It suggests that telmisartan may play an anti-inflammatory effect by blocking the IKCa1 potassium channels of T lymphocyte activation.

[Voltage-dependent potassium channel and calcium-activated potassium channel current changes of peripheral blood T-lymphocytes from hypertensive patients in Xinjiang Kazakh].

OBJECTIVE: To observe the current changes of voltage-dependent potassium channel (Kv1.3 potassium channel) and calcium-activated potassium channel (IKCa1 potassium channel) in peripheral blood T-lymphocyte derived from hypertensive patients of Xinjiang Kazakh. METHODS: Twenty randomly selected untreated Kazakh hypertensive patients and 20 Kazakh healthy subjects from Xinjiang were included in this study. T-lymphocytes were isolated from peripheral blood with magnetic cell sorting, the whole-cell currents of Kv1.3 and IKCa1 potassium channels were recorded with patch-clamp technique. RESULTS: (1) The current density of Kv1.3 potassium channel was significantly higher in the hypertensive group [(280 ± 74) pA/pF (n = 39)] than that in the control group [(179 ± 51) pA/pF (n = 38), P < 0.01], while the membrane capacitance was similar between the two groups. (2) The current density of IKCa1 potassium channel was also significantly higher in the hypertensive group [(198 ± 44) pA/pF (n = 28)] than that in the control group [(124 ± 43) pA/pF (n = 26), P < 0.01], while the membrane capacitance was also similar between the two groups. CONCLUSIONS: The T-lymphocytes Kv1.3 potassium channel and IKCa1 potassium channel current densities are higher in hypertensive patients in Xinjiang Kazakh suggesting a potential role of Kv1.3 and IKCa1 potassium channels activation in the pathophysiology of hypertension.

Aqueous Chloride-Ion Battery within a Neutral Electrolyte Based on a CoFe-Cl Layered Double Hydroxide Anode.

Aqueous chloride-ion batteries (ACIBs) with environmental friendliness and high safety hold great potential to fulfill the green energy demand for ocean desalination. Herein, for the first time, a composite consisting of Cl--intercalated CoFe layered double hydroxides (CoFe-Cl-LDH) cross-linked with CNTs (CoFe-Cl-LDH/CNT) is synthesized and demonstrated to be a novel high-performance anode for ACIBs in a neutral NaCl aqueous solution. While exhibiting a high initial capacity of ∼190 mAh g-1 at 200 mA g-1, CoFe-Cl-LDH/CNT is capable of delivering a reversible capacity of ∼125 mAh g-1 after 200 cycles. At a high current density of 400 mA g-1, it still holds a capacity of ∼120 mAh g-1. The excellent Cl- storage performance can be contributed to the unique topochemical transformation feature that reverses intercalation/deintercalation of Cl- along with valence changes of Co2+/Co3+ and Fe2+/Fe3+ during charge/discharge and the improved electronic conductivity by hybridizing with CNTs. It is interesting that the invertible insertion/extraction of interlayer H2O was discovered, which could be beneficial to the capacity after cycles to a certain extent. The Cl--intercalated LDH material declared in this work shows its feasibility on Cl- capture/release in aqueous anion-type batteries and provides a new opportunity for future development of ACIBs or aqueous desalination technology.

An MgAl layered double hydroxide as a new transition metal-free anode for lithium-ion batteries.

A carbonate intercalated magnesium aluminum layered double hydroxide is used as an anode material for lithium-ion batteries, displaying a maximum discharge specific capacity of 814 mA h g-1 at 200 mA g-1 in this work through utilizing the valence variation of Mg and the conversion between LiOH and LiH/Li2O.

MgAl Saponite as a Transition-Metal-Free Anode Material for Lithium-Ion Batteries.

Transition-metal compounds (oxides, sulfides, hydroxides, etc.) as lithium-ion battery (LIB) anodes usually show extraordinary capacity larger than the theoretical value due to the transformation of LiOH into Li2O/LiH. However, there has rarely been a report relaying the transformation of LiOH into Li2O/LiH as the main reaction for LIBs, due to the strong alkalinity of LiOH leading to battery deterioration. In this work, layered silicate MgAl saponite (MA-SAP) is applied as a -OH donor to generate LiOH as the anode material of LIBs for the first time. The MA-SAP maintains a layered structure during the (dis)charging process and has zero-strain characteristic on the (001) crystal plane. In the discharging process, Mg, Al, and Si in the saponite sheets become electron-rich, while the active hydroxyl groups escape from the sheets and combine with lithium ions to form LiOH in the "caves" on sheets, and the LiOH continues to decompose into Li2O and LiH. Consequently, the MA-SAP delivers a maximum capacity of 536 mA h·g-1 at 200 mA·g-1 with a good high-current discharging ability of 155 mA h·g-1 after 1000 cycles under 1 A·g-1. Considering its extremely low cost and completely nontoxic characteristics, MA-SAP has great application prospects in energy storage. In addition, this work has an enlightening effect on the development of new anodes based on extraordinary mechanisms.

A Zero-Strain Insertion Cathode Material for Room-Temperature Fluoride-Ion Batteries.

A fluoride-ion battery (FIB) is a novel type of energy storage system that has a higher volumetric energy density and low cost. However, the high working temperature (>150 °C) and unsatisfactory cycling performance of cathode materials are not favorable for their practical application. Herein, fluoride ion-intercalated CoFe layered double hydroxide (LDH) (CoFe-F LDH) was prepared by a facile co-precipitation approach combined with ion-exchange. The CoFe-F LDH shows a reversible capacity of ∼50 mAh g-1 after 100 cycles at room temperature. Although there is still a big gap between FIBs and lithium-ion batteries, the CoFe-F LDH is superior to most cathode materials for FIBs. Another important advantage of CoFe-F LDH FIBs is that they can work at room temperature, which has been rarely achieved in previous reports. The superior performance stems from the unique topochemical transformation property and small volume change (∼0.82%) of LDH in electrochemical cycles. Such a tiny volume change makes LDH a zero-strain cathode material for FIBs. The 2D diffusion pathways and weak interaction between fluoride ions and host layers facilitate the de/intercalation of fluoride ions, accompanied by the chemical state changes of Co2+/Co3+ and Fe2+/Fe3+ couples. First-principles calculations also reveal a low F- diffusion barrier during the cyclic process. These findings expand the application field of LDH materials and propose a novel avenue for the designs of cathode materials toward room-temperature FIBs.

Acute toxicity and safety profile evolution of aqueous extracts of Sanren decoction using in vivo models.

ETHNOPHARMACOLOGICAL RELEVANCE: Sanren decoction (SRD) is a well-known traditional Chinese medicine prescription containing eight kinds of materials. SRD has been used mainly in China for more than 200 years for the treatment of respiratory disorders that co-occur with a bad fever after midday. AIM OF THE STUDY: To evaluate the acute and 28-day subacute toxicity of an aqueous extract of SRD using in vivo methods. MATERIALS AND METHODS: To determine acute toxicity, SRD was administered by gavage at a dosage of 58.5 g/kg/day to male and female mice for 7 days. To determine subacute toxicity, SRD was administered at 3.3, 6.5, or 13 g/kg/day to male and female rats for 28 days. The general behavior, body weight, biochemical and hematological parameters, organ coefficients and pathological morphology of the treated animals were analyzed. RESULTS: Neither acute nor subacute concentrations of SRD caused significant changes in the body weights, general behavior, hematology and biochemical parameters, organ weights, or histopathological appearances of the liver, kidney, spleen, brain, lung or heart in mice or rats. CONCLUSIONS: The administration of SRD can be considered safe within the conditions of this study.

[Development and evaluation of a novel method for rapid screening of Pichia pastoris strains capable of efficiently expressing recombinant proteins].

Pichia pastoris is one of the most widely used recombinant protein expression systems. In this study, a novel method for rapid screening of P. pastoris strains capable of efficiently expressing recombinant proteins was developed. Firstly, the ability to express recombinant proteins of the modified strain GS115-E in which a functional Sec63-EGFP (Enhanced green fluorescent protein) fusion protein replaced the endogenous endoplasmic reticulum transmembrane protein Sec63 was tested. Next, the plasmids carrying different copy numbers of phytase (phy) gene or xylanase (xyn) gene were transformed into GS115-E to obtain recombinant strains with different expression levels of phytase or xylanase, and the expression levels of EGFP and recombinant proteins in different strains were tested. Finally, a flow cytometer sorter was used to separate a mixture of cells with different phytase expression levels into sub-populations according to green fluorescence intensity. A good linear correlation was found between the fluorescence intensities of EGFP and the expression levels of the recombinant proteins in the recombinant strains (0.8<|R|<1). By using the flow cytometer, high-yielding P. pastoris cells were efficiently screened from a mixture of cells. The expression level of phytase of the selected high-fluorescence strains was 4.09 times higher than that of the low-fluorescence strains after 120 h of methanol induction. By detecting the EGFP fluorescence intensity instead of detecting the expression level and activity of the recombinant proteins in the recombinant strains, the method developed by the present study possesses the greatly improved performance of convenience and versatility in screening high-yielding P. pastoris strains. Combining the method with high-throughput screening instruments and technologies, such as flow cytometer and droplet microfluidics, the speed and throughput of this method will be further increased. This method will provide a simple and rapid approach for screening and obtaining P. pastoris with high abilities to express recombinant proteins.

Evaluation of a cell model expressing ßKlotho for screening FGF21 analogues.

ßKlotho as the major role is a necessary auxiliary protein when fibroblast growth factor 21 (FGF21) binds FGF21 receptors (FGFR) for activating intracellular signaling pathways that ultimately generate biological effects. To achieve the aim of high throughput screening of FGF21 analogues, we established 3T3-L1-ßKlotho cells that could stably express ßklotho protein. The glucose uptake, expression of GLUT1 mRNA and activation of FGF signaling molecules ERK1/2 phosphorylation were detected by GOD-POD assay, real-time PCR analysis and western blotting assay in 3T3-L1-ßKlotho cells and 3T3-L1 adipocytes, respectively. The results showed that FGF21 increased glucose uptake significantly in a dose-dependent and time-dependent manner in 3T3-L1-ßKlotho cells. 3T3-L1-ßKlotho cells stimulated with FGF21 up-regulated the transcriptional levels of GLUT1 mRNA obviously. FGF21 activated the FGF signaling molecules ERK1/2 in 3T3-L1-ßKlotho cells. In addition, the same results were obtained in 3T3-L1 adipocytes. Furthermore, FGF21-stimulated elevation of glucose uptake, GLUT1 mRNA transcription and the phosphorylation of ERK1/2 were dramatically attenuated by pretreatment of cells with FGFR specific inhibitor SU5402 in 3T3-L1-ßKlotho cells. This study demonstrated that the cell model could be applied to high throughput screen FGF21 analogues.

[Progresses in inheritance of genes for avian eggshell color].

Eggshell has three colors: white, blue and brown. Chicken and duck eggs with blue eggshell have superior market for its better appearance, delicious taste, abundant nutrition and higher eggshell thickness and strength compared to those with white eggshell. However, error was often made when breeding blue-eggshell chicken or duck lines based on phenotypes. Studies on the forming and controlling mechanism of eggshell color had important theoretic and practical value. This review mainly discussed the types of eggshell color, its pigment composition and synthesis. Inheritance and heritability, genetic model, the number of genes, and the dominant-recessive relationship between genes for eggshell color were also reviewed. Information described in this review is useful for understanding the forming mechanism of eggshell color.

Recycling of a selectable marker with a self-excisable plasmid in Pichia pastoris.

Pichia pastoris is a widely used heterologous protein production workhorse. However, with its multiple genetic modifications to solve bottlenecks for heterologous protein productivity, P. pastoris lacks selectable markers. Existing selectable marker recycling plasmids have drawbacks (e.g., slow growth and conditional lethality). Here, zeocin-resistance marker recycling vectors were constructed using the Cre/loxP recombination system. The vectors were used to (i) knock in heterologous phytase, xylanase and lipase expression cassettes, (ii) increase the phytase, xylanase and lipase gene copy number to 13, 5, and 5, respectively, with vector introduction and (iii) engineer the secretion pathway by co-overexpressing secretion helper factors (Sly1p and Sec1p) without introducing selectable markers, giving a phytase field of 0.833 g/L. The vectors allow selectable marker recycling and would be a useful tool to engineer P. pastoris for high heterologous protein productivity.

Anti-human fibroblast growth factor-21 monoclonal antibody preparation, characterization and analysis of in vitro bioactivity.

Human fibroblast growth factor 21 (hFGF-21) is involved in numerous metabolic processes and elevated hFGF-21 levels are associated with many metabolic diseases. However, the role hFGF-21 serves in the metabolic system is not fully understood. A humanized anti-hFGF-21 monoclonal antibody (mAb) would provide a novel method for further investigations into the role hFGF-21 serves in the metabolic system and related diseases, which may reveal therapeutic targets for future treatment of these diseases. The present study aimed to prepare an anti-hFGF-21 mAb, followed by identification of its characteristics and bioactivity in vitro. The results of the present study identified that the anti-hFGF-21 mAb (clone 2D8) produced had good specificity, had an immunoglobulin isotype of IgG2b and a titer of 1:1.024×106. hFGF-21 was screened for epitopes using fluorescence-activated cell sorting, which revealed a specific 15 amino acid sequence (YQSEAHGLPLHLPGN) that the anti-hFGF-21 mAb recognized. In vitro bioactivity of anti-hFGF-21 was determined using a glucose uptake assay and by measuring the expression of glucose transporter 1 (GLUT1) messenger RNA (mRNA) in 3T3-L1 adipocytes. This revealed that hFGF-21-dependent glucose uptake and GLUT1 mRNA expression were negatively correlated with increasing levels of the anti-hFGF-21 mAb tested, and that hFGF-21 activity could be overcome by increasing concentrations of the mAb, demonstrating that the mAb has hFGF-21-neutralizing activity in vitro.

Inhibitory effects of telmisartan on culture and proliferation of and Kv1.3 potassium channel expression in peripheral blood CD4+ T lymphocytes from Xinjiang Kazakh patients with hypertension.

INTRODUCTION: Activation of T lymphocytes, for which potassium channels are essential, is involved in the development of hypertension. In this study, we explored the inhibitory effects of telmisartan on the culture and proliferation of and Kv1.3 potassium channel expression in peripheral blood CD4+ T lymphocytes derived from Xinjiang Kazakh patients with hypertension. METHODS: CD4+ T-cell samples from hypertensive Kazakh patients and healthy Kazakh people were divided into healthy control, case control, telmisartan, and 4-aminopytidine groups. Changes in the expression levels of interleukin (IL)-6 and IL-17 in the blood of the healthy control and case control subjects were detected by enzyme-linked immunosorbent assay. Peripheral blood CD4+ T lymphocytes were first activated and proliferated in vitro and then incubated for 0, 24, and 48 h under various treatment conditions. Thereafter, changes in CD4+ T-lymphocytic proliferation were determined using Cell Counting Kit-8 and microscope photography. Changes in messenger RNA (mRNA) and protein expression of the Kv1.3 potassium channel in CD4+ T lymphocytes were detected using real-time quantitative polymerase chain reaction and Western blots, respectively. RESULTS: The IL-6 and IL-17 expression levels were significantly higher in the blood of the hypertensive Kazakh patients than in the healthy Kazakh people. Telmisartan inhibited T-lymphocytic proliferation, as well as the mRNA and protein expression of the Kv1.3 potassium channel in CD4+ T lymphocytes, and the inhibitory effects were time-dependent, with the strongest inhibition observed after 48 h and significantly weaker inhibition observed after 24 h of treatment. CONCLUSIONS: Telmisartan may potentially regulate hypertensive inflammatory responses by inhibiting T-lymphocytic proliferation and Kv1.3 potassium channel expression in CD4+ T lymphocytes.